Heterologous Expression and Engineering of the Nitrogenase Cofactor Biosynthesis Scaffold NifEN

Solomon, Joseph B.; Lee, Chi Chung; Jasniewski, Andrew J.; Rasekh, Mahtab F.; Ribbe, Markus W.; Hu, Yilin

doi:10.1002/anie.201916598

Heterologous Expression and Engineering of the Nitrogenase Cofactor Biosynthesis Scaffold NifEN

Journal Article · Tue Feb 04 23:00:00 EST 2020 · Angewandte Chemie (International Edition)

DOI:https://doi.org/10.1002/anie.201916598· OSTI ID:1803013

Solomon, Joseph B. ^[1]; Lee, Chi Chung ^[2]; Jasniewski, Andrew J. ^[2]; Rasekh, Mahtab F. ^[3]; ^[3]; Hu, Yilin ^[2]

Univ. of California, Irvine, CA (United States). Dept. of Molecular Biology & Biochemistry. Dept. Chemistry; OSTI
Univ. of California, Irvine, CA (United States). Dept. of Molecular Biology & Biochemistry
Univ. of California, Irvine, CA (United States). Dept. of Molecular Biology & Biochemistry. Dept. Chemistry

NifEN plays a crucial role in the biosynthesis of nitrogenase, catalyzing the final step of cofactor maturation prior to delivering the cofactor to NifDK, the catalytic component of nitrogenase. The difficulty in expressing NifEN, a complex, heteromultimeric metalloprotein sharing structural/functional homology with NifDK, is a major challenge in the heterologous expression of nitrogenase. Herein, we report the expression and engineering of Azotobacter vinelandii NifEN in Escherichia coli. Biochemical and spectroscopic analyses demonstrate the integrity of the heterologously expressed NifEN in composition and functionality and, additionally, the ability of an engineered NifEN variant to mimic NifDK in retaining the matured cofactor at an analogous cofactor-binding site. This is an important step toward piecing together a viable pathway for the heterologous expression of nitrogenase and identifying variants for the mechanistic investigation of this enzyme.

View Accepted Manuscript (DOE)

Research Organization:: Univ. of California, Irvine, CA (United States)

Sponsoring Organization:: USDOE Office of Science (SC), Basic Energy Sciences (BES)

Grant/Contract Number:: SC0016510

OSTI ID:: 1803013

Alternate ID(s):: OSTI ID: 1602596

Journal Information:: Angewandte Chemie (International Edition), Journal Name: Angewandte Chemie (International Edition) Journal Issue: 17 Vol. 59; ISSN 1433-7851

Publisher:: WileyCopyright Statement

Country of Publication:: United States

Language:: English

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